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目的观察兔骨髓基质细胞来源的细胞凝集态在体外钙结节形成过程的形态变化。方法通过全骨髓培养方法分离培养兔骨髓基质细胞,并在培养瓶中进行扩增,使用成骨诱导液作为培养基;再通过四环素标记、碱性磷酸酶染色、茜素红染色、扫描电镜及透射电镜方法,研究细胞凝集态在钙结节形成过程中的形态变化。结果荧光显微镜下观察可见,四环素标记的钙结节发出金黄色荧光;光镜下见结节中的细胞凝集态与周边铺壁细胞相比呈碱性磷酸酶及茜素红的强阳性反应;扫描电镜下见结节中的凝集态细胞均呈球状生长并分泌基质;透射电镜下见细胞中基质小泡丰富。结论骨髓来源的基质细胞在体外形成的钙结节需要三维空间状态下细胞的高度凝集。
OBJECTIVE: To observe the morphology of rabbit bone marrow stromal cell-derived aggregates in vitro during calcium formation. Methods Rabbit bone marrow stromal cells were isolated and cultured by whole bone marrow culture and expanded in culture flasks. Osteogenic medium was used as culture medium. Tetracycline labeling, alkaline phosphatase staining, alizarin red staining and scanning electron microscopy Transmission electron microscopy was used to investigate the morphological changes of the cell aggregates during the formation of calcium nodules. Results Fluorescence microscopy showed that the tetracycline-labeled calcium nodules fluoresced golden yellow. Strong positive reactions of alkaline phosphatase and alizarin red were observed in the condensed state of nodules in the nodules compared with the surrounding parietal cells under the light microscope. Under the scanning electron microscope, the agglutinated cells in the nodules all showed spherical growth and secreted matrix; under the transmission electron microscope, the matrix vesicles were abundant. Conclusion Bone marrow-derived stromal cells require highly aggregated cells in three-dimensional space in vitro.